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CNPY2 / Protein canopy homolog 2 · IHC design guide
Design Immunohistochemistry for CNPY2
Plan CNPY2 IHC in paraffin sections using the reported cytoplasmic tissue pattern (HPA tissue IHC). Compare glandular or placental trophoblastic staining with appropriate controls, mindful that the tissue profile may reflect protein from more than one gene (HPA tissue IHC).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
CNPY2 Immunohistochemistry Experimental Design Guide
Expected localisation, validated protocols, controls and antibodies — the at-a-glance facts below, then the full design guide.
Must know before staining
| Expected localisation | Cytoplasmic staining in tissue (HPA tissue IHC) | |
| Staining pattern | Glandular and trophoblastic cells show cytoplasmic staining (HPA tissue IHC) | |
| Antigen retrieval | Citrate pH 6.0 HIER, heat-mediated (datasheet A10020) | |
| Positive control | Cervix+4 more · see all | |
| Negative control | Oral mucosa |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Keep fixation consistent across compared paraffin sections. (selected-SKU IHC image A10020) | |
| Caveat | Staining may reflect protein from more than one gene (HPA tissue IHC) | |
| Regulation | Highest in adult placenta, liver and pancreas (UniProt) | |
| Isoform / epitope | Two isoforms; signal peptide 1–20 is cleaved; check epitope (UniProt) |
Section 1
Recommended CNPY2 IHC & IF Protocols
The catalog antibody’s IHC-P protocol is accompanied by published CNPY2 protocols for mouse liver, rat myocardium, and mouse tissues (PMC8306219; PMC6443342; PMC4230931).
| Sample | Paraffin-embedded mouse kidney tissue; fixative not specified (datasheet A10020) |
| Fixation | Image fixative and duration unreported (datasheet A10020); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat retrieval: Citrate pH 6.0 (datasheet A10020); 20 min, 95–100 °C (standard) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% normal serum of the secondary host, 30 min, room temperature (standard) |
| Primary antibody | Rabbit anti-CNPY2, 1:50-1:200 (datasheet A10020) |
| Primary incubation | Overnight at 4 °C (standard) |
| Detection | HRP-polymer secondary, DAB chromogen 5–10 min (standard) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | CNPY2-positive staining in glandular cells of cervix (HPA tissue IHC: High). HPA tissue profile: Ubiquitous cytoplasmic expression. No signal in the no-primary control. |
💡Decision noteStart with citrate pH 6.0 heat retrieval (datasheet A10020); it was also used in mouse liver and gave the most sensitive detection among tested buffers in mouse tissues (PMC8306219; PMC4230931).
Section 2
What Is the Expected CNPY2 Staining Pattern?
Expect predominantly cytoplasmic CNPY2 staining in tissue sections, particularly in glandular cells and placental trophoblastic cells (HPA tissue IHC: ubiquitous cytoplasmic expression; High in these cells). UniProt places CNPY2 in the endoplasmic reticulum and reports no transmembrane segment (UniProt Q9Y2B0: subcellular location and topology). HPA rates its tissue IHC pattern Supported, while cautioning that the staining may include protein from more than one gene (HPA tissue IHC: reliability).
| Cytoplasmic staining in placental trophoblastic cells or glandular cells of cervix, epididymis, fallopian tube, or thyroid. | This fits the reported high staining in those cell populations (HPA tissue IHC: High). Compare the signal within the identified cells; a positive section need not show uniform staining across all cell types (HPA tissue IHC: cell-specific levels). |
| Predominantly nuclear staining, with little or no cytoplasmic signal. | A nuclear-only pattern conflicts with the reported tissue pattern and protein location (HPA tissue IHC: cytoplasmic; UniProt Q9Y2B0: endoplasmic reticulum). Treat it as suspect and check the detection control before assigning it to CNPY2 (general IHC practice). |
| Strong staining in oral mucosal squamous epithelial cells. | That cell population is reported as not detected (HPA tissue IHC: oral mucosa). Consider nonspecific binding or endogenous detection activity; use an appropriate reagent control and assess staining by cell type (general IHC practice). |
| Similar haze or precipitate across cells, extracellular spaces, and the section edge. | A diffuse pattern is difficult to reconcile with cell-associated cytoplasmic staining (HPA tissue IHC: profile). Check reagent controls, blocking, washing, and chromogen development for background (general IHC practice). |
| No stain in a section containing a reported high-staining cell population. | An absent result in those cells conflicts with the reference pattern (HPA tissue IHC: High in listed glandular and trophoblastic cells). First establish that the staining run worked using a known-positive section; a single negative section does not establish absent CNPY2 expression (general IHC practice). |
💡Expected CNPY2 appearanceCall a positive result when identifiable glandular or trophoblastic cells show clear cytoplasmic chromogen above background, especially in HPA high-staining sites; isolated nuclear stain or broad cell-free haze is suspect (HPA tissue IHC: cytoplasmic, High; general IHC practice).
| Cell population and tissue | HPA reports High staining in selected glandular cells and placental trophoblastic cells, Medium in adipocytes, and Low in several other populations (HPA tissue IHC: cell-level entries). Judge the specified cells rather than the whole section. |
| Intracellular location | UniProt assigns CNPY2 to the endoplasmic reticulum and reports no transmembrane segment (UniProt Q9Y2B0: location, topology). HPA describes the tissue IHC pattern as cytoplasmic, without resolving an organelle-scale pattern (HPA tissue IHC: profile). |
| Protein processing | The annotated signal peptide spans residues 1–20, followed by a 21–182 chain (UniProt Q9Y2B0: processing). No antibody epitope is supplied, so this record cannot predict whether processing changes recognition in the section. |
| Antibody evidence | Two listed antibodies have Supported IHC status; one has Approved ICC status (HPA antibodies: HPA038465, HPA038466). HPA cautions that tissue staining may include protein from more than one gene, so morphology alone cannot establish molecular specificity (HPA tissue IHC: reliability description). |
| Isoforms | Two isoforms are listed (UniProt Q9Y2B0: isoforms 1 and 2). Their presence alone gives no section-level staining prediction without an epitope map or isoform-specific IHC evidence. |
| IF/ICC Q: What location should I expect? | A: HPA reports approved cytosol localization in ICC-IF, with images from A-431, U-251MG, and U2OS (HPA subcellular: main location, image list). That observation is assay-specific; UniProt separately annotates endoplasmic-reticulum location (UniProt Q9Y2B0). |
| Situation | Likely cause | Next action |
|---|---|---|
| No signal in a reported high-staining cell population. | The stain may have failed, or the relevant cells may be absent from the examined area (HPA tissue IHC: cell-level High entries; general IHC practice). | Confirm the cell population on the counterstain and run a known-positive section in the same staining run; review primary-antibody and detection steps (general IHC practice). |
| Weak stain where a reference cell population is marked Low. | Low staining is reported for several populations, including parathyroid glandular cells and cardiomyocytes (HPA tissue IHC: Low entries). | Compare with a reported High population before calling the run unsuccessful; score comparable cell types with the same background threshold (HPA tissue IHC: High and Low; general IHC practice). |
| Nuclear-only staining dominates the section. | The compartment conflicts with cytoplasmic tissue staining and annotated endoplasmic-reticulum location (HPA tissue IHC: profile; UniProt Q9Y2B0). | Inspect a reagent control and a reported positive cell population; reassess specificity before scoring the nuclear signal as CNPY2 (general IHC practice). |
| Brown signal appears in many unrelated structures. | Nonspecific reagent binding or endogenous enzyme activity can produce chromogenic background (general IHC practice). | Use the relevant reagent control; review blocking, washes, and enzyme-blocking steps appropriate to the detection chemistry (general IHC practice). |
| Oral mucosal squamous cells stain strongly. | Those cells are reported as not detected, and HPA cautions that staining may include protein from more than one gene (HPA tissue IHC: oral mucosa, reliability description). | Check morphology and a control lacking primary antibody; compare staining with a reported high-staining population before interpreting it as specific (general IHC practice). |
| A result differs between IHC and IF/ICC. | HPA reports cytoplasmic tissue IHC and approved cytosol localization in ICC-IF, while UniProt annotates endoplasmic-reticulum location (HPA tissue IHC; HPA subcellular; UniProt Q9Y2B0). | Interpret each assay against its own reported pattern and controls; do not infer a tissue-section organelle pattern from the ICC-IF summary alone (general IHC/IF practice). |
Experiment evidence
IHC & IF images showing CNPY2 staining
A10020 · IHCImmunohistochemistry of paraffin-embedded mouse kidney using CNPY2 Rabbit pAb at dilution of 1:100 .Perform high pressure antigen retrieval
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000257727-CNPY2/tissue/cerebral+cortex#imid_9985537
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000257727-CNPY2/tissue/colon#imid_9985455
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000257727-CNPY2/tissue/liver#imid_9985464
HPA · KidneyHuman Protein Atlas IHC, Kidney. Source: https://www.proteinatlas.org/ENSG00000257727-CNPY2/tissue/kidney#imid_9985465
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA038465; green = CNPY2, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA038465; green = CNPY2, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for CNPY2 IHC & IF
🧪Run placenta first: trophoblastic cells should stain (HPA: High in placental trophoblastic cells). Run oral mucosa as the negative tissue (HPA: Not detected in oral-mucosa squamous epithelial cells); on the placenta slide, assess non-trophoblastic cells for background staining, but do not assume they are target-negative (standard IHC practice).
Positive control tissue: Cervix (Glandular cells, HPA High)
Negative control tissue: Oral mucosa (HPA Not detected)
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show CNPY2 in A-431, U-251MG, U2OS, with annotated localisation: Cytosol (approved) (HPA subcellular).
Technical controls: Include no-primary (secondary-only) and isotype-matched rabbit IgG controls for the rabbit polyclonal catalog antibody, plus a CNPY2 knockout or validated peptide-block control (selected SKU caption: rabbit pAb; standard IHC practice). Check endogenous peroxidase background on the placenta slide before interpreting chromogenic signal (standard IHC practice).
⚠️Feasibility: A target-specific fixation window or fixation effect is unreported, and the selected A10020 paraffin-section mouse-kidney caption does not state the fixative (selected SKU caption). That caption uses high-pressure retrieval in 10 mM citrate buffer at pH 6.0 and an antibody dilution of 1:100; it does not establish retrieval requirements for placenta or whether frozen sections are easier (selected SKU caption). IF/ICC images exist for A-431, U-251MG and U2OS, but the supplied evidence does not establish that IF is easier; assess placental background separately from specific trophoblastic staining (HPA subcellular; HPA: High in placental trophoblastic cells; standard IHC practice).
HPA tissue IHC evidence for CNPY2
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Supported — High consistency between antibody staining and RNA expression data. Caution, targets protein from more than one gene.). Rows are taken directly from the HPA tissue chart — click any row's HPA link to view the source.
Positive expression · recommended positive controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Cervix | Glandular cells | High | Protein (IHC) | HPA → |
| Epididymis | Glandular cells | High | Protein (IHC) | HPA → |
| Fallopian tube | Glandular cells | High | Protein (IHC) | HPA → |
| Placenta | Trophoblastic cells | High | Protein (IHC) | HPA → |
| Thyroid gland | Glandular cells | High | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Oral mucosa | Squamous epithelial cells | Not detected | Protein (IHC) | HPA → |
Section 3
Advanced CNPY2 IHC Tips
Troubleshoot CNPY2 staining in paraffin sections by checking retrieval, compartment, controls and scoring before interpreting chromogenic signal.
What retrieval should I try first when CNPY2 staining is weak?
Use heat-mediated retrieval in 10 mM citrate buffer, pH 6.0, before staining paraffin sections (datasheet A10020). The selected mouse kidney image used high-pressure retrieval with this buffer and a 1:100 antibody dilution; it does not establish an optimal heating time for other tissues (caption A10020). If signal remains weak, compare a small retrieval time series while holding antibody dilution and chromogenic development constant, and watch for tissue damage (standard IHC practice). Record staining intensity alongside morphology and a no-primary control so stronger background is not mistaken for improved antigen exposure (standard IHC practice).
How should I troubleshoot variable CNPY2 staining after fixation?
CNPY2-specific fixation sensitivity is unknown: the selected paraffin-section caption does not state a fixative, so its staining cannot establish a preferred fixation condition (caption A10020). Record the fixative and exposure time for each specimen, then compare sections processed and stained together with the same retrieval and antibody dilution (standard IHC practice). Check morphology and background as well as signal, because overprocessing and uneven fixation can complicate chromogenic interpretation (standard IHC practice). Do not attribute a fixation effect to CNPY2’s endoplasmic-reticulum annotation, signal peptide or modified residue without a controlled fixation comparison (UniProt Q9Y2B0; standard IHC practice).
Where should convincing CNPY2 chromogenic staining appear?
Start by assessing cytoplasmic staining: tissue IHC reports ubiquitous cytoplasmic expression, while the subcellular IF assessment assigns CNPY2 to the cytosol (HPA tissue IHC; HPA subcellular). UniProt also annotates the endoplasmic reticulum and no transmembrane segment, so record diffuse and perinuclear cytoplasmic patterns separately rather than forcing one pattern (UniProt Q9Y2B0). Compare cellular staining with adjacent unstained structures and a no-primary control under identical chromogen development (standard IHC practice). Predominantly nuclear or extracellular deposits need additional scrutiny before being scored as CNPY2, given the reported cytoplasmic localisation (HPA tissue IHC; standard IHC practice).
Could isoforms or epitope accessibility explain discordant staining?
CNPY2 has 2 annotated isoforms, a signal peptide at residues 1–20, and a mature chain spanning 21–182 (UniProt Q9Y2B0). Check the antibody’s documented immunogen against those regions and the isoforms under study before treating discordant IHC results as biological differences (UniProt Q9Y2B0; standard IHC practice). The supplied caption does not locate this antibody’s epitope, so staining cannot establish which isoform or processed form it detects (caption A10020). If retrieval changes signal, compare matched sections and an independent antibody with a documented, distinct epitope before assigning an isoform-specific explanation (standard IHC practice).
How can I compare CNPY2 IF with the chromogenic IHC pattern?
Use IF as a complementary localisation check, keeping its controls and scoring separate from this antibody’s paraffin-section IHC result (caption A10020; standard IF practice). For multiplexing, pair CNPY2 with a validated trophoblastic-cell marker in placenta, where tissue IHC reports high staining in trophoblastic cells (HPA tissue IHC). Choose spectrally separated fluorophores and inspect an unstained section, because tissue autofluorescence can mimic a weak channel (standard IF practice). Determine whether the antibody recognises an accessible cytosolic epitope or one requiring entry into the endoplasmic-reticulum lumen before choosing permeabilisation; the epitope position is not supplied here (UniProt Q9Y2B0; standard IF practice).
What controls help distinguish diffuse CNPY2 signal from background?
CNPY2 is reported as broadly cytoplasmic in tissue IHC, so diffuse brown staining alone is insufficient to establish specificity (HPA tissue IHC; standard IHC practice). Run no-primary and secondary-only controls, then inspect whether precipitate or endogenous enzyme activity persists when the primary antibody is omitted (standard IHC practice). For peroxidase-based detection, include a peroxidase block and compare chromogen development times on matched sections; these are general workflow controls (standard IHC practice). If background remains high, titrate around the caption’s 1:100 dilution and judge cellular contrast against morphology, rather than selecting the darkest section (caption A10020; standard IHC practice).
How should I score CNPY2 across heterogeneous tissue sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and tissue area before scoring, then use the same inclusion rules, counterstain and chromogen development across specimens (standard IHC practice). Report either the percentage of positive cells with intensity categories or an H-score from 0–300, with thresholds set before comparing groups (standard IHC practice). Where positive cells occur in discrete regions, positive-cell density per mm² can supplement those measures if the analysed area is reported (standard IHC practice). Normalise to the eligible cells or viable tissue area in each section, and report compartments separately when diffuse and perinuclear cytoplasmic patterns differ (HPA tissue IHC; UniProt Q9Y2B0; standard IHC practice).
When should an apparent CNPY2-positive focus be rejected?
A plausible positive focus should show cellular cytoplasmic staining in intact tissue, consistent with the reported IHC pattern and cytosolic localisation assessment (HPA tissue IHC; HPA subcellular). High staining in placental trophoblastic cells or selected glandular cells provides a tissue reference, while oral-mucosa squamous epithelial cells are reported as not detected (HPA tissue IHC). Treat edge-restricted signal, necrotic debris, extracellular precipitate or colour present in the no-primary control as potential artefact, including endogenous enzyme activity (standard IHC practice). Because the tissue IHC reliability note cautions that staining may target protein from more than one gene, resolve surprising patterns with an independent specificity control before assigning them to CNPY2 (HPA tissue IHC; standard IHC practice).
Boster reagents
Best CNPY2 / Protein canopy homolog 2 IHC Antibodies
One anti-CNPY2 antibody has real IHC data from paraffin-embedded mouse kidney (A10020 IHC image caption). Human, Mouse, and Rat reactivity is listed (catalog: reactivity).
Real IHC data
Anti-Protein canopy homolog 2 CNPY2 Antibody
Cat # A10020
A10020 is the only card and shows IHC staining of paraffin-embedded mouse kidney (A10020 IHC image caption). Its listed reactivity is Human, Mouse, and Rat; no IF image is supplied (catalog: reactivity, IF image alts).
Which to pick: Choose A10020 for paraffin-section tissue IHC: its rabbit polyclonal image uses 1:100 dilution after high-pressure retrieval in 10 mM citrate buffer at pH 6.0; the fixative is unreported (A10020 IHC image caption). For cross-species work, it lists Human, Mouse, and Rat reactivity, while the displayed IHC evidence is from mouse kidney (catalog: reactivity; A10020 IHC image caption). No IF/ICC option is supported by this catalog entry (catalog: applications, IF image alts, IF dilution).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry Q9Y2B0 (CNPY2_HUMAN, Protein canopy homolog 2).
- Human Protein Atlas. CNPY2 tissue IHC expression (reliability: Supported).
- Human Protein Atlas. CNPY2 subcellular location (ICC-IF): Localized to the cytosol..
- Human Protein Atlas. CNPY2 antibody validation summary (2 antibodies).
- Canopy Homolog 2 as a Novel Molecular Target in Hepatocarcinogenesis. Cancers 2021 — PMC8306219.
- Protective effect of Barbaloin in a rat model of myocardial ischemia reperfusion injury through the regulation of the CNPY2‑PERK pathway. International journal of molecular medicine 2019 — PMC6443342.
- Expression of CNPY2 in mouse tissues: quantification and localization. PloS one 2014 — PMC4230931.
- CNPY2 inhibits MYLIP-mediated AR protein degradation in prostate cancer cells. Oncotarget 2018 — PMC5915145.
- PubMed PMID:10072769 — UniProt-cited evidence.
- PubMed PMID:12826659 — UniProt-cited evidence.
- PubMed PMID:12975309 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for CNPY2 (Protein canopy homolog 2). Expected localisation Cytoplasmic staining in tissue (HPA tissue IHC). Antigen retrieval: Citrate pH 6.0 HIER, heat-mediated (datasheet A10020). Validated IHC antibody: A10020 at 1:50-1:200 (the product's own IHC validation image is shown). Controls (HPA): positive Cervix (Glandular cells); negative Oral mucosa. IHC workflow; specimen preparation unspecified. Protocol steps: 1. Sample: Paraffin-embedded mouse kidney tissue; fixative not specified (datasheet A10020); 2. Fixation: Image fixative and duration unreported (datasheet A10020); verify before use.; 3. Sectioning: 4–5 µm sections on charged slides (standard); 4. Deparaffinisation: Xylene, graded ethanol series to water (standard); 5. Antigen retrieval: Heat retrieval: Citrate pH 6.0 (datasheet A10020); 20 min, 95–100 °C (standard); 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% normal serum of the secondary host, 30 min , room temperature (standard); 8. Primary antibody: Rabbit anti-CNPY2, 1:50-1:200 (datasheet A10020); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Keep fixation consistent across compared paraffin sections. (selected-SKU IHC image A10020); Caveat: Staining may reflect protein from more than one gene (HPA tissue IHC); Isoform / epitope: Two isoforms; signal peptide 1–20 is cleaved; check epitope (UniProt); Regulation: Highest in adult placenta, liver and pancreas (UniProt). Evidence: UniProt Q9Y2B0, HPA tissue IHC · Supported, HPA ICC-IF · cytosol., Datasheet A10020, PMC8306219, PMC6443342. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/CNPY2/immunohistochemistry-ihc-guide.